LOCUS       sequence-159431-        7140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7140)
  AUTHORS   Lee-Fruman KK, Kuo CJ, Lippincott J, Terada N, Blenis J
  TITLE     Characterization of S6K2, a novel kinase homologous to S6K1.
  JOURNAL   Oncogene. 1999 Sep 9. 18(36):5108-14.
  PUBMED    10490847
REFERENCE   2  (bases 1 to 7140)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Oncogene. 
            1999 Sep 9. 18(36):5108-14."
COMMENT     Sequence Label: pcDNA3 S6K2 E388 D3E
FEATURES             Location/Qualifiers
     source          1..7140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             909..935
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2693..2711)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2694..2711)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2731..2748)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2737..2961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3007..3435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3094..3113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3304..3325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3444..3464)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3449..3778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3629..3764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3691..3710
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3845..4639
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3899..3918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4509..4528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4813..4934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4850..4869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4904..4923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4983..4999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..4999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4996..5018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5007..5023
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5031..5037,5038..5055,5056..5061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5076..5097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5214..5231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5385..5973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5465..5484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6144..6935,6936..7004))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6767..6786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7005..7109)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttcgatcg
      901 tcgacatgta cccatacgat gttccagatt acgcttctag aggatccgcg gccgtgtttg
      961 atttggattt ggagacggag gaaggcagcg agggcgaggg cgagccagag ctcagccccg
     1021 cggacgcatg tccccttgcc gagttgaggg cagctggcct agagcctgtg ggacactatg
     1081 aagaggtgga gctgactgag accagcgtga acgttggccc agagcgcatc gggccccact
     1141 gctttgagct gctgcgtgtg ctgggcaagg ggggctatgg caaggtgttc caggtgcgaa
     1201 aggtgcaagg caccaacttg ggcaaaatat atgccatgaa agtcctaagg aaggccaaaa
     1261 ttgtgcgcaa tgccaaggac acagcacaca cacgggctga gcggaacatt ctagagtcag
     1321 tgaagcaccc ctttattgtg gaactggcct atgccttcca gactggtggc aaactctacc
     1381 tcatccttga gtgcctcagt ggtggcgagc tcttcacgca tctggagcga gagggcatct
     1441 tcctggaaga tacggcctgc ttctacctgg ctgagatcac gctggccctg ggccatctcc
     1501 actcccaggg catcatctac cgggacctca agcccgagaa catcatgctc agcagccagg
     1561 gccacatcaa actgaccgac tttggactct gcaaggagtc tatccatgag ggcgccgtca
     1621 ctcacacctt ctgcggcacc attgagtaca tggcccctga gattctggtg cgcagtggcc
     1681 acaaccgggc tgtggactgg tggagcctgg gggccctgat gtacgacatg ctcactggat
     1741 cgccgccctt taccgcagag aaccggaaga aaaccatgga taagatcatc aggggcaagc
     1801 tggcactgcc cccctacctc accccagatg cccgggacct tgtcaaaaag tttctgaaac
     1861 ggaatcccag ccagcggatt gggggtggcc caggggatgc tgctgatgtg cagagacatc
     1921 cctttttccg gcacatgaat tgggacgacc ttctggcctg gcgtgtggac ccccctttca
     1981 ggccctgtct gcagtcagag gaggacgtga gccagtttga tacccgcttc acacggcaga
     2041 cgccggtgga cagtcctgat gacacagccc tcagcgagag tgccaaccag gccttcctgg
     2101 gcttcgaata cgtggcgccg tctgtcctgg acagcatcaa ggagggcttc tccttccagc
     2161 ccaagcttcg cgaccccagg cgcctcaaca gtgacccccg ggtccccgtc gaccccctca
     2221 agttctcccc ttttgagggg tttcggccca gccccagcct gccggagccc acggagctac
     2281 ctctacctcc actcctgcca ccgccgccgc cctcgaccac cgcccctctc cccatccgtc
     2341 ccccctcagg gaccaagaag tccaagaggg gccgtgggcg tccagggcgc taggaagccg
     2401 ggtgggggtg agggtagccc ttgagccctg tccctgcggc tgtgagagca gcaggaccct
     2461 gggccagttc cagagacctg ggggtgtgtc tgggggtggg gtgtgagtgc gtatgaaagt
     2521 gtgtgtctgc tggggcagct gtgcccctga atcatgggca cggagggccg cccgccacac
     2581 cccgcgctca actgctcccg tggaagatta aagggctgaa tcatctcgtg ccgaattctg
     2641 cagatatcca tcacactggc ggccgctcga gcatgcatct agagggccct attctatagt
     2701 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     2761 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2821 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2881 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2941 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3001 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3061 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3421 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3781 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4021 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggctg accgctatca
     4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4921 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5041 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
